CNP對Ach引起的糖尿病大鼠胃平滑肌收縮增強的影響及IGF-1的干預效應
發(fā)布時間：2018-11-02 19:04
【摘要】：目的：本實驗通過觀察C型鈉尿肽、乙酰膽堿、硝普鈉等藥物對STZ誘導的糖尿病模型大鼠胃自發(fā)性收縮活動的作用以及IGF-1對糖尿病胃動力障礙的治療效果,進一步探討糖尿病胃動力障礙的發(fā)病機理,以期為尋找防治糖尿病胃動力障礙的有效方案提供理論依據(jù)。方法：Wistar雄性大鼠30只(250g±20g),隨機分為正常對照組、模型組、IGF-1治療組各10只。STZ腹腔注射制備糖尿病大鼠模型,糖尿病造模成功后第10周對IGF-1治療組行腹腔注射IGF-1(1500ng/kg)治療2周,均于造模12周后進行如下實驗。測量體質(zhì)量、血糖、尿量；通過多道生理信號記錄系統(tǒng)記錄各組大鼠胃竇平滑肌自發(fā)性收縮活動；對各組大鼠胃竇平滑肌組織進行HE染色以觀察其形態(tài)結構；采用ELISA測定血清CNP的含量；利用RT-PCR觀察胃組織中CNP mRNA的表達；免疫組織化學方法觀察各組大鼠胃平滑肌組織中NPR-A和NPR-B的分布；利用電鏡觀察胃竇平滑肌細胞內(nèi)超微結構；利用Western bolt觀察胃平滑肌組織中GRP78的表達。結果:1.模型組及IGF-1治療組大鼠血糖濃度明顯高于正常對照組大鼠(n=6,P0.01),體重明顯低于正常對照組大鼠(n=6,P0.05),尿量明顯多于正常對照組大鼠(n=6,P0.01),差異均有統(tǒng)計學意義;2.HE染色示模型組及IGF-1治療組大鼠胃竇平滑肌變薄；IGF-1治療組與模型組間無顯著性差異。3.模型組及IGF-1治療組大鼠胃收縮張力、頻率較正常組明顯減低,對乙酰膽堿敏感性降低,對CNP、SNP敏感性增加,差異均有統(tǒng)計學意義(n=6,p0.05)。4.免疫組織化學結果顯示模型組及IGF-1治療組大鼠與正常對照組相比胃竇平滑肌組織NPR-B表達明顯增多,差異具有統(tǒng)計學意義(n=9,p0.05),各組大鼠胃平滑肌中NPR-A表達無顯著性差異。5.IGF-1治療組大鼠胃肌條收縮、胃平滑肌形態(tài)學改變、胃平滑肌組織NPR-B、 NPR-A較模型組無顯著性差異(n=6,P0.05)。6. ELISA結果顯示模型組及IGF-1治療組大鼠血清中的CNP較正常對照組無顯著性差異(n=6,P0.05)。7. RT-PCR結果顯示模型組及工GF-1治療組大鼠胃竇平滑肌組織中的CNPmRNA表達較正常組明顯增多,差異具有統(tǒng)計學意義(n=6,P0.05)。8.電鏡下可見,與對照組相比,模型組及IGF-1治療組平滑肌細胞內(nèi)質(zhì)網(wǎng)明顯腫脹、空泡化,線粒體極消失,但IGF-1治療組較模型組明顯改善(n=6,P0.05)。9. Western結果顯示IGF-1治療組GRP78表達明顯低于模型組,差異具有統(tǒng)計學意義(n=6,P0.05)。結論：STZ誘導的糖尿病大鼠發(fā)病12周后,胃竇平滑肌自發(fā)性收縮活動明顯異常,收縮頻率減慢、振幅明顯下降、收縮節(jié)律紊亂；內(nèi)源性CNP參與糖尿病胃動力障礙大鼠胃收縮的調(diào)節(jié)；胃竇平滑肌NP-NPR-B-cGMP和NO-sGC-cGMP信號轉導途徑的活性明顯提高；IGF-1治療對糖尿病胃動力障礙大鼠胃收縮活動無明顯改善作用,但是可以阻止或改善內(nèi)質(zhì)網(wǎng)應激作用。
[Abstract]:Objective: to observe the effect of type C natriuretic peptide, acetylcholine and sodium nitroprusside on gastric spontaneous contraction induced by STZ in diabetic rats and the therapeutic effect of IGF-1 on diabetic gastric motility. To explore the pathogenesis of diabetic gastric motility disorder in order to provide theoretical basis for finding an effective scheme for prevention and treatment of diabetic gastric motility disorder. Methods: thirty Wistar male rats (250g 鹵20g) were randomly divided into normal control group, model group and IGF-1 treatment group. Diabetic rats were induced by intraperitoneal injection of STZ. The IGF-1 group was treated with intraperitoneal injection of IGF-1 (1500ng/kg) for 2 weeks at the 10th week after successful diabetic modeling. The following experiments were carried out 12 weeks later. Body mass, blood glucose and urine volume were measured. Spontaneous contraction of gastric antral smooth muscle was recorded by multi-channel physiological signal recording system. The antral smooth muscle tissue of each group was stained with HE to observe its morphological structure. The content of serum CNP was measured by ELISA, the expression of CNP mRNA in gastric tissue was observed by RT-PCR, the distribution of NPR-A and NPR-B in gastric smooth muscle tissue was observed by immunohistochemical method. The ultrastructure of gastric antral smooth muscle cells and the expression of GRP78 in gastric smooth muscle tissue were observed by electron microscope and Western bolt respectively. The result is 1: 1. The blood glucose concentration in the model group and the IGF-1 group was significantly higher than that in the normal control group (n = 6, P 0.01), and the body weight was significantly lower than that in the normal control group (P 0.05), and the urine volume was significantly higher than that in the normal control group (n = 6, P 0.01). The differences were statistically significant. 2.HE staining showed that gastric antrum smooth muscle thinned in model group and IGF-1 group. There was no significant difference between IGF-1 treatment group and model group. The gastric contractile tension and frequency in model group and IGF-1 group were significantly lower than those in normal group, and the sensitivity to acetylcholine was decreased, and the sensitivity to CNP,SNP was increased, with significant difference (nong6, p0.05). 4. The results of immunohistochemistry showed that the expression of NPR-B in the antral smooth muscle tissue of the model group and the IGF-1 group was significantly higher than that of the normal control group, and the difference was statistically significant. There was no significant difference in the expression of NPR-A in the gastric smooth muscle of rats in each group. The gastric muscle strips contracted and the gastric smooth muscle morphologically changed in the 5.IGF-1 treatment group. There was no significant difference in NPR-B, NPR-A in the gastric smooth muscle tissue compared with the model group (n = 6, P < 0.05). P0.05). 6. ELISA results showed that there was no significant difference in serum CNP between the model group and the IGF-1 treatment group compared with the normal control group (7. 05). The results of RT-PCR showed that the expression of CNPmRNA in gastric antral smooth muscle tissue in model group and GF-1 group was significantly higher than that in normal group (nti6P 0.05). Compared with the control group, the endoplasmic reticulum of smooth muscle cells in the model group and IGF-1 treatment group was obviously swollen, vacuolated and mitochondria disappeared, but the IGF-1 group was significantly improved compared with the model group (P 0.05). Western results showed that the expression of GRP78 in IGF-1 group was significantly lower than that in model group (P 0.05). Conclusion: the spontaneous contractile activity of gastric antral smooth muscle in diabetic rats induced by STZ was obviously abnormal, the contractile frequency slowed down, the amplitude decreased, and the contraction rhythm was disturbed 12 weeks after onset. Endogenous CNP was involved in the regulation of gastric contraction in diabetic rats with gastric motility disorder, and the activity of NP-NPR-B-cGMP and NO-sGC-cGMP signal transduction pathway in gastric antral smooth muscle was significantly increased. IGF-1 treatment did not significantly improve gastric contractile activity in diabetic rats with gastric motility disorder, but could prevent or improve endoplasmic reticulum stress.
【學位授予單位】：延邊大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R587.2
本文編號：2306684
[Abstract]:Objective: to observe the effect of type C natriuretic peptide, acetylcholine and sodium nitroprusside on gastric spontaneous contraction induced by STZ in diabetic rats and the therapeutic effect of IGF-1 on diabetic gastric motility. To explore the pathogenesis of diabetic gastric motility disorder in order to provide theoretical basis for finding an effective scheme for prevention and treatment of diabetic gastric motility disorder. Methods: thirty Wistar male rats (250g 鹵20g) were randomly divided into normal control group, model group and IGF-1 treatment group. Diabetic rats were induced by intraperitoneal injection of STZ. The IGF-1 group was treated with intraperitoneal injection of IGF-1 (1500ng/kg) for 2 weeks at the 10th week after successful diabetic modeling. The following experiments were carried out 12 weeks later. Body mass, blood glucose and urine volume were measured. Spontaneous contraction of gastric antral smooth muscle was recorded by multi-channel physiological signal recording system. The antral smooth muscle tissue of each group was stained with HE to observe its morphological structure. The content of serum CNP was measured by ELISA, the expression of CNP mRNA in gastric tissue was observed by RT-PCR, the distribution of NPR-A and NPR-B in gastric smooth muscle tissue was observed by immunohistochemical method. The ultrastructure of gastric antral smooth muscle cells and the expression of GRP78 in gastric smooth muscle tissue were observed by electron microscope and Western bolt respectively. The result is 1: 1. The blood glucose concentration in the model group and the IGF-1 group was significantly higher than that in the normal control group (n = 6, P 0.01), and the body weight was significantly lower than that in the normal control group (P 0.05), and the urine volume was significantly higher than that in the normal control group (n = 6, P 0.01). The differences were statistically significant. 2.HE staining showed that gastric antrum smooth muscle thinned in model group and IGF-1 group. There was no significant difference between IGF-1 treatment group and model group. The gastric contractile tension and frequency in model group and IGF-1 group were significantly lower than those in normal group, and the sensitivity to acetylcholine was decreased, and the sensitivity to CNP,SNP was increased, with significant difference (nong6, p0.05). 4. The results of immunohistochemistry showed that the expression of NPR-B in the antral smooth muscle tissue of the model group and the IGF-1 group was significantly higher than that of the normal control group, and the difference was statistically significant. There was no significant difference in the expression of NPR-A in the gastric smooth muscle of rats in each group. The gastric muscle strips contracted and the gastric smooth muscle morphologically changed in the 5.IGF-1 treatment group. There was no significant difference in NPR-B, NPR-A in the gastric smooth muscle tissue compared with the model group (n = 6, P < 0.05). P0.05). 6. ELISA results showed that there was no significant difference in serum CNP between the model group and the IGF-1 treatment group compared with the normal control group (7. 05). The results of RT-PCR showed that the expression of CNPmRNA in gastric antral smooth muscle tissue in model group and GF-1 group was significantly higher than that in normal group (nti6P 0.05). Compared with the control group, the endoplasmic reticulum of smooth muscle cells in the model group and IGF-1 treatment group was obviously swollen, vacuolated and mitochondria disappeared, but the IGF-1 group was significantly improved compared with the model group (P 0.05). Western results showed that the expression of GRP78 in IGF-1 group was significantly lower than that in model group (P 0.05). Conclusion: the spontaneous contractile activity of gastric antral smooth muscle in diabetic rats induced by STZ was obviously abnormal, the contractile frequency slowed down, the amplitude decreased, and the contraction rhythm was disturbed 12 weeks after onset. Endogenous CNP was involved in the regulation of gastric contraction in diabetic rats with gastric motility disorder, and the activity of NP-NPR-B-cGMP and NO-sGC-cGMP signal transduction pathway in gastric antral smooth muscle was significantly increased. IGF-1 treatment did not significantly improve gastric contractile activity in diabetic rats with gastric motility disorder, but could prevent or improve endoplasmic reticulum stress.
【學位授予單位】：延邊大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R587.2
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